title: A surface plasmon resonance-based biosensor with full-length BACE1 in a reconstituted membrane.
abstract: A surface plasmon resonance (SPR) biosensor-based assay for membrane-embedded full-length BACE1 (β-site amyloid precursor protein cleaving enzyme 1), a drug target for Alzheimer's disease, has been developed. It allows the analysis of interactions with the protein in its natural lipid membrane environment. The enzyme was captured via an antibody recognizing a C-terminal His6 tag, after which a lipid membrane was reconstituted on the chip using a brain lipid extract. The interaction between the enzyme and several inhibitors confirmed that the surface was functional. It had slightly different interaction characteristics as compared with a reference surface with immobilized ectodomain BACE1 but had the same inhibitor characteristic pH effect. The possibility of studying interactions with BACE1 under more physiological conditions than assays using truncated enzyme or conditions dictated by high enzyme activity is expected to increase our understanding of the role of BACE1 in Alzheimer's disease and contribute to the discovery of clinically efficient BACE1 inhibitors. The strategy exploited in the current study can be adapted to other membrane-bound drug targets by selecting suitable capture antibodies and lipid mixtures for membrane reconstitution.
authors.affiliation: Department of Biochemistry and Organic Chemistry, Uppsala University, Sweden.
authors.forename: Tony
authors.initials: T
authors.lastname: Christopeit
authors.forename: Gun
authors.initials: G
authors.lastname: Stenberg
authors.forename: Thomas
authors.initials: T
authors.lastname: Gossas
authors.forename: Susanne
authors.initials: S
authors.lastname: Nyström
authors.forename: Vera
authors.initials: V
authors.lastname: Baraznenok
authors.forename: Erik
authors.initials: E
authors.lastname: Lindström
authors.forename: U Helena
authors.initials: UH
authors.lastname: Danielson
mesh_terms: D000544:Alzheimer Disease / Q000188:drug therapy / Q000201:enzymology; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000037:antagonists & inhibitors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008051:Lipid Bilayers / Q000378:metabolism; D008958:Models, Molecular; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D008051:Lipid Bilayers; D053829:Amyloid Precursor Protein Secretases; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human; D002118:Calcium